The Capillary Technology group (Project 5) will explore the application of capillary electrophoresis and microfluidics to DNA sequencing and DNA based diagnostics. A prototype diagnostic instrument has been constructed which is capable of performing PCR amplifications in a small diameter tube, purifying the reactions and efficiently loading the products onto a capillary electrophoresis instrument for analysis. The entire procedure is automated in a simple fashion. The complete reaction, purification and analysis is complete in under 30 minutes. The extension of this system to DNA sequencing is the primary goal of this proposal. Applications in genotyping using microsatellite repeat polymorphisms and PCR diagnostics will also be pursued. Improvements to the basic instrument will be made in the areas of throughput, sensitivity, economy and utility. A multi (TM) capillary version of the instrument will be constructed in the latter part of the grant. Our group will also continue to examine the applicability of the technology we developed for reloading capillary gels to slab gel systems. We will work to reduce the effort and cost of our present methods, while increasing the number of times such gels can be used beyond our current limit of four consecutive runs.